459 Peptide YY (PYY)3–36 modulates thyrotropin secretion in rats K J Oliveira, G S M Paula, R H Costa-e-Sousa, L L Souza, D C Moraes, F H Curty and C C Pazos-Moura Laborato´rio de Endocrinologia Molecular, Instituto de Biofı´sica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21949-900, Brazil (Requests for offprints should be addressed to C C Pazos-Moura; Email: [email protected]) Abstract Peptide YY (PYY)3-36 is a gut-derived hormone, with a However, in fasted rats, PYY3-36 at both doses elicited a proposed role in central mediation of postprandial satiety signals, significant rise (approximately twofold increase, P!0.05) in as well as in long-term energy balance. In addition, recently,the serum TSH observed 15 min after the hormone injection. ability of the hormone to regulate gonadotropin secretion, PYY3-36 treatment did not modify significantly serum T4,T3,or acting at pituitary and at hypothalamus has been reported. Here, leptin. Therefore, in the present paper, we have demonstrated we examined PYY3-36 effects on thyrotropin (TSH) secretion, that the gut hormone PYY3-36 acts directly on the pituitary both in vitro and in vivo.PYY3-36-incubated rat pituitary glands gland to inhibit TSH release, and in the fasting situation, in vivo, showed a dose-dependent decrease in TSH release, with 44 and when serum PYY3-36 is reduced, the activity of thyroid axis is K K 62% reduction at 10 8 and 10 6 M(P!0.05 and P!0.001 reduced as well. In such a situation, systemically injected respectively), and no alteration in TSH response to thyrotropin- PYY3-36 was able to acutely activate the thyrotrope axis, releasing hormone. In vivo,PYY3-36 i.p. single injection in the suggesting a new role for PYY3-36 as a regulator of the doses of 3 or 30 mg/kg body weight, administered to rats fed ad hypothalamic–pituitary–thyroid axis. libitum, was not able to change serum TSH after 15 or 30 min. Journal of Endocrinology (2006) 191, 459–463 Introduction (Keire et al. 2000). Experimental evidence suggests that circulating PYY3-36 inhibits appetite by acting directly on Peptide YY (PYY)3-36 is a gut-derived hormone, produced the arcuate nucleus via the Y2 receptor, a presynaptic by endocrine L cells lining the distal small bowel and colon, inhibitory autoreceptor (Batterham et al. 2002, Talsania et al. released postprandially in proportion to the calories ingested 2005). NPY is a potent stimulator of food intake and PYY3-36 (Adrian et al. 1985). Peripheral administration of PYY3-36 in was able to inhibit the electrical activity of arcuate NPY rodents or humans has been reported to induce a marked neurons as well as to reduce the expression of the NPY mRNA inhibition of food intake (Batterham et al. 2002, 2003). (Batterham et al. 2002, Challisa et al. 2003). In addition, NPY PYY3-36 inhibits food intake at plasma concentrations that are originating in the hypothalamic arcuate nucleus exerts a within the normal physiological range seen in man after meals profound inhibitory effect on the thyroid axis via effects on (Batterham et al. 2002, 2003) and therefore, a role for the hypophysiotropic thyrotropin-releasing hormone (TRH) hormone in central mediation of postprandial satiety signals neurons. Chronic intracerebroventricular administration of has been proposed. Moreover, long-term administration has NPY to normally fed rats resulted in reduction of circulating been demonstrated to induce a reduction in body weight levels of thyroid hormones with inappropriately normal or low (Batterham et al. 2002, 2003) although this is still a matter of thyrotropin (TSH), and suppression of proTRH mRNA in the controversy (Tscho¨p et al. 2004). hypothalamic paraventricular nucleus (PVN; Fekete et al. In addition, recent studies suggested that PYY3-36 may act 2001). NPYeffects were reproduced by NPY-Y1and NPY-Y5 as a neuroendocrine regulator. It has been demonstrated in analogs injected into the third cerebral ventricle, suggesting prepubertal rats that PYY3-36 stimulated prolactin, luteinizing that both the receptors mediate NPY suppression of the hormone (LH), and FSH secretion acting directly at pituitary hypothalamic–pituitary–thyroid axis (Fekete et al. 2002). gland, although it seems to have an inhibitory action at the During fasting, as a means of conserving energy, the hypothalamic level (Aguilar et al. 2004, Fernandez-Fernandez hypothalamic–pituitary–thyroid axis is suppressed and the et al. 2005). Therefore, similar to other hormones involved in activation of arcuate NPY neurons is a major component of energy homeostasis, PYY3-36 is also able to influence the the regulatory mechanism that causes a decline in proTRH reproductive axis. However, there is no information mRNA in PVN, reducing circulating levels of TSH and concerning the thyrotrope axis. thyroid hormones. Fasting is associated with reduced serum PYY3-36 is a member of the neuropeptide Y (NPY) family concentrations of PYY3-36 (Tov a r et al. 2004, Chan et al. and it is an agonist of receptor subtypes NPY-Y2and NPY-Y5 2005), and peripherally injected PYY3-36 partly reversed the Journal of Endocrinology (2006) 191, 459–463 DOI: 10.1677/joe.1.06784 0022–0795/06/0191–459 q 2006 Society for Endocrinology Printed in Great Britain Online version via http://www.endocrinology-journals.org Downloaded from Bioscientifica.com at 09/27/2021 07:08:43PM via free access 460 K J OLIVEIRA and others . PYY3-36 and rat TSH secretion fasting-induced c-Fos expression in arcuate nucleus neurons Quantification of TSH of mice (Riediger et al. 2004), suggesting a role for the TSH concentration in the serum and in the incubation peptide in fasting adaptation. medium was measured by specific RIA, employing reagents Therefore, considering NPYeffects on thyroid axis, and the supplied by the National Institute of Diabetes, Digestive and presence of Y5 receptors, and the mRNA encoding Y2 and Kidney Diseases, National Hormone & Peptide Program Y5 receptors in pituitary gland (Parker et al. 2000, (NIDDK–NHPP) (Torrance, CA, USA), as previously Fernandez-Fernandez et al. 2005), we tested the hypothesis described (Chard 1987, Ortiga-Carvalho et al. 1996), and that PYY may act directly at the pituitary to modulate 3-36 was expressed in terms of the reference preparation 3 (RP3). TSH secretion. In addition, we also investigated whether Within-assay variation was 5.7%. Samples of the same PYY , injected systemically, may modify the thyrotrope 3-36 experiment were measured within the same assay. Minimum axis in fed and fasting rats. assay detection was 0.52 ng/ml. Materials and Methods Quantification of serum concentrations of T4,T3, and leptin Animals Serum T4 and T3 were detected by RIA (MP Biomedicals Inc, Irvine, CA, USA). Detection limits were: 1 mg/dl for T Adult male Wistar rats, weighing 250–300 g, were kept under 4 and 25 ng/dl for T . Serum leptin was measured using a controlled lighting (12 h light:12 h darkness cycle, lights on at 3 specific rat RIA by LINCO Research (St Charles, MO, 0700 h) and controlled temperature (23G1 8C). All experi- USA). Minimum detectable level was 0.5 ng/ml. Intraassay mental protocols were approved by our institutional animal variation was less than 7% for all hormone measurements and care committee. all the samples were run within the same assay. In vitro experiments Statistical analysis Ad libitum fed rats were killed by decapitation, and their Data are reported as meansGS.E.M. One-way ANOVA anterior pituitaries were quickly dissected out for in vitro followed by a Student–Newman–Keuls multiple comparisons testing as described before (Rettori et al. 1989, Ortiga- test was employed for the assessment of significance of data. Carvalho et al. 2002) Each hemi-pituitary was immediately Serum TSH was analyzed after logarithmic transformation transferred to a flask containing 1 ml Krebs–Ringer (Zar 1996). Differences were considered to be significant at bicarbonate medium (pH 7.4) at 37 8C in an atmosphere of P!0.05. 95% O2/5% CO2 in a Dubnoff metabolic shaker. After a 30-min preincubation period, medium was changed to 1 ml medium alone (control) or medium containing PYY3-36 (Bachem California, Inc., Torrance, CA, USA) to a final K K K Results concentration of 10 10,10 8,or10 6 M. At the end of a 2 h incubation period, an aliquot was removed for TSH In vitro experiments measurement. In another set of experiments, after 2 h incubation, a small aliquot was removed for TSH measure- PYY3-36-incubated hemi-pituitary glands showed, after 2 h ment and TRH (Bachem California, Inc.) was added to a final incubation, a dose-dependent decrease in TSH release, K K concentration of 50 nM in all tubes. The incubation was statistically significant at 10 8 M(P!0.05) and 10 6 M continued for 30 min to determine the TRH-stimulated (P!0.001), with a reduction of 44 and 62% respectively TSH release in the absence or presence of the different (Fig. 1). However, the TSH response to TRH was not concentrations of PYY3-36. significantly different among groups, presenting an increment around four times above the basal levels (TSH before TRH) in all groups. In vivo experiments Ad libitum fed rats were divided into three groups that In vivo experiments received a single i.p. injection of 3 or 30 mg/kg body weight (BW) PYY3-36 or 0.2 ml saline vehicle (control group). The i.p. administration of 3 or 30 mg/kg BW of PYY3-36 to Another set of rats was fasted for 3 days before they received a ad libitum fed rats induced no changes in serum TSH either single i.p.